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Development and evaluation of a competitive enzyme-linked immunosorbent assay using a monoclonal antibody for diagnosis of severe fever with thrombocytopenia syndrome virus in bovine sera

View Article: PubMed Central - PubMed

ABSTRACT

Severe fever with thrombocytopenia syndrome (SFTS) caused by the SFTS virus (SFTSV), a phlebovirus in the family Bunyaviridae, is an emerging tick-borne infectious disease that impacts humans. This disease manifests as a decreased blood cell count and multi-organ failure, with a case-fatality rate of more than 12% in China. Because vaccines or antiviral drugs for the treatment of this disease are not available, monitoring the SFTS circulation in animals and controlling the tick-mammal cycle are important for preventing SFTS. Monoclonal antibodies against the recombinant nucleoprotein of SFTSV were generated to develop a competitive enzyme-linked immunosorbent assay (cELISA) for the detection of antibodies against SFTSV infection in cattle. The specificity and sensitivity of cELISA was assessed by comparing the results of this assay to those of an immunofluorescence assay (IFA). The results of the cELISA using 416 field bovine serum samples and laboratory-immunized positive sera showed 98.1% consistency with those of the IFA. The cELISA used in this study did not show cross-reactivity with antisera against other viral cattle diseases. The cELISA presented in this study can be applied to detect antibodies against SFTSV in cattle.

No MeSH data available.


The percent inhibition (PI) value and optical density (OD) of laboratory immunized bovine sera in a competitive enzyme-linked immunosorbent assay (cELISA). Serum samples collected from two immunized cattle and one negative control cattle were diluted from 1/2 to 1/300 and tested for (A) PI value and (B) OD value (at 450–630 nm) in cELISA. Error bars represent the standard error of the mean of independent experiments repeated at least three times.
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Figure 4: The percent inhibition (PI) value and optical density (OD) of laboratory immunized bovine sera in a competitive enzyme-linked immunosorbent assay (cELISA). Serum samples collected from two immunized cattle and one negative control cattle were diluted from 1/2 to 1/300 and tested for (A) PI value and (B) OD value (at 450–630 nm) in cELISA. Error bars represent the standard error of the mean of independent experiments repeated at least three times.

Mentions: The immunized cattle serum samples were tested for the cELISA. Both 1 : 2 and 1 : 5 dilutions of the positive serum samples showed higher PI values than the negative serum. However, a 1 : 5 dilution of the positive control sera yielded a large difference in the optical density (OD) between the positive and negative serum samples (Fig. 4). The repeatability of this finding was confirmed by three independent experiments.


Development and evaluation of a competitive enzyme-linked immunosorbent assay using a monoclonal antibody for diagnosis of severe fever with thrombocytopenia syndrome virus in bovine sera
The percent inhibition (PI) value and optical density (OD) of laboratory immunized bovine sera in a competitive enzyme-linked immunosorbent assay (cELISA). Serum samples collected from two immunized cattle and one negative control cattle were diluted from 1/2 to 1/300 and tested for (A) PI value and (B) OD value (at 450–630 nm) in cELISA. Error bars represent the standard error of the mean of independent experiments repeated at least three times.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037297&req=5

Figure 4: The percent inhibition (PI) value and optical density (OD) of laboratory immunized bovine sera in a competitive enzyme-linked immunosorbent assay (cELISA). Serum samples collected from two immunized cattle and one negative control cattle were diluted from 1/2 to 1/300 and tested for (A) PI value and (B) OD value (at 450–630 nm) in cELISA. Error bars represent the standard error of the mean of independent experiments repeated at least three times.
Mentions: The immunized cattle serum samples were tested for the cELISA. Both 1 : 2 and 1 : 5 dilutions of the positive serum samples showed higher PI values than the negative serum. However, a 1 : 5 dilution of the positive control sera yielded a large difference in the optical density (OD) between the positive and negative serum samples (Fig. 4). The repeatability of this finding was confirmed by three independent experiments.

View Article: PubMed Central - PubMed

ABSTRACT

Severe fever with thrombocytopenia syndrome (SFTS) caused by the SFTS virus (SFTSV), a phlebovirus in the family Bunyaviridae, is an emerging tick-borne infectious disease that impacts humans. This disease manifests as a decreased blood cell count and multi-organ failure, with a case-fatality rate of more than 12% in China. Because vaccines or antiviral drugs for the treatment of this disease are not available, monitoring the SFTS circulation in animals and controlling the tick-mammal cycle are important for preventing SFTS. Monoclonal antibodies against the recombinant nucleoprotein of SFTSV were generated to develop a competitive enzyme-linked immunosorbent assay (cELISA) for the detection of antibodies against SFTSV infection in cattle. The specificity and sensitivity of cELISA was assessed by comparing the results of this assay to those of an immunofluorescence assay (IFA). The results of the cELISA using 416 field bovine serum samples and laboratory-immunized positive sera showed 98.1% consistency with those of the IFA. The cELISA used in this study did not show cross-reactivity with antisera against other viral cattle diseases. The cELISA presented in this study can be applied to detect antibodies against SFTSV in cattle.

No MeSH data available.